Cell viability and toxicity service

service eca viability new2

PROCOMCURE Biotech offers a broad selection of cell-based and biochemical screening services for the analysis of your compounds, as characterization of mode of action and functionality are crucial for the development of a viable drug.

The cell-based methods allow you to generate a cellular profile of your compound using endpoints such as NADH-, ATP-, and cAMP-content and protease activity.

The read-out is either based on a luminescent or a fluorescent reaction. We established a flexible portfolio of methods, which can be customized to your need (see also pathway reporter screening).

 

 

 

Assay
Choose your assay and get suggestions for complementary experiments
Cell line
Choose the appropriate cell line
Setup
Choose the concentration range of your compound and incubation time or ask our staff for suggestions
Results
The data is delivered in tables and graphs
Compound
Please provide us with data about solubility and concentration of your test compound

 

Service Catalog

ApplicationMarkerDetection How it worksNotes
cell viability, cell toxicity, proliferationATPluminescenceThe amount of ATP is measured by an ATP-dependent luciferase reaction, where luciferin is metabolized and results in emission of light. available for eukaryotic and prokaryotic cells
cell viability, cell toxicity, proliferationNADHfluorescenceMetabolically active cells are able to convert an indicator dye (by a reduction reaction), which can be detected by fluorometric measurement. flexible incubation times possible
cell viability, cell toxicityintracellular proteasefluorescenceA protease acts as marker for living cells and converts a membrane-permeable peptide into a fluorescent product and hence generates a signal which is proportional to cell viability.can be used for normalization (well to well and plate to plate differences)
cell toxicity, membrane damageDNAfluorescenceDNA is released from dead cells and bound by the dye, which is non-toxic and non-membranepermeable. The fluorescent signal increases proportional with the released DNA, as equivalent to the number of dead cells. can be used in addition to ATP cell viability assay
cell toxicity, membrane damageextracellular proteaseluminescenceBecause dead cells lose their membrane integrity, a specific protease is released into the medium. This enzyme cleaves a peptide-substrate, which can then serve as substrate in a luciferase reaction. The emissioned light is a measure of the number of damaged cells. cells stay intact, normalization possible
apoptosiscaspase-3 and -7luminescenceThe activity of the effector caspases -3 and -7 is a value which shows the number of apoptotic cells as a result of compound treating. The caspases cleave a peptide that can then serve as substrate in a luciferase reaction.
GPCR signaling pathwaycAMP contentluminescenceWhen a compound interacts with G-protein coupled receptors (GPCRs) linked with adenylate cyclase, the intracellular level of cAMP is altered. The cAMP acts as a stimulant for the protein kinase A (PKA) in the reagent. The active kinase metabolizes ATP that decreases the simultaneous luciferase reaction and the emitted light. That means, the lower the luminescence counts, the higher the cAMP level.
chymotrypsin-, trypsin- and caspase-like activity of the proteasomesubstrate conversionluminescenceAll three activities of the proteasome can be measured combined in a single experiment or each of them separately. The substrate is added and processed by the proteasome to serve as reagent for a luciferase reaction. Proteasome inhibitors are of great interest in the development of new cancer therapies.
cell viability, cell toxicityMTTcolorimetricMTT substrate is taken up by treated cells and - depending on mitochondrial integrity / activity as overall measure of cell viability - reduced to an impermeable (intracellularly accumulating) purple formazan product. Lysis of cells causes a liberation of the substrate which can be quantified via colorimetric procedures.
cardio toxicityhERG channel proteinfluorescence polarizationA high-affinity fluorescent ligand (a so-called tracer) is herein displaced from the hERG channel protein (included in a membrane fraction) by compounds that bind to it, as has been validated with a panel of known blockers. Results demonstrate a high correlation with those obtained from patch clamp techniques.

 

Discuss your cell viability/toxicity project...